Donnini Sandra, Solito Raffaella, Giachetti Antonio, Granger Harris J, Ziche Marina, Morbidelli Lucia
Pharmacology Section, Department of Molecular Biology, Via A. Moro 2, 53100 Siena, Italy.
J Pharmacol Exp Ther. 2006 Nov;319(2):515-22. doi: 10.1124/jpet.106.108803. Epub 2006 Jul 25.
The beneficial effect exerted by angiotensin-converting enzyme inhibitors (ACEI) on vascular endothelium has been attributed to restoration of endothelial cell survival properties and improvement of angiogenesis. Fibroblast growth factor (FGF)-2 is an angiogenic factor for the microvascular endothelium, which tonically promotes endothelial cell growth and survival through an autocrine/paracrine mechanism. Here, we formulate the hypothesis that FGF-2 might contribute to the prosurvival/proangiogenic effect of ACEI. We investigated zofenoprilat and, in selected experiments, lisinopril, as representatives of ACEI. These compounds induced formation of pseudocapillaries in vessel fragments isolated from porcine coronary and human umbilical arteries by increasing endothelial cell growth up to 5-fold. Angiogenesis was abolished by inhibitors of nitric-oxide synthase (NOS) pathway and by anti-FGF-2 antibodies. Consistently, in cultured coronary endothelial cells (CVECs), ACEI up-regulated endothelial NOS (eNOS) and FGF-2 and induced mitogen-activated protein kinase extracellular signal-regulated kinase 1/2 activation. The overexpression of eNOS/FGF-2 produced, at the functional level, enhanced cell proliferation and migration, the latter effect being dose-dependent and maximal at 0.1 microM zofenoprilat. The importance of FGF-2 for the acquisition of the angiogenic phenotype elicited by ACEI was clearly demonstrated by the impairment of endothelial functions following transfection of CVECs with small interference RNA for FGF-2. Moreover, FGF-2 silencing greatly affected the nuclear translocation of the FGF receptor (FGFR)-1, highlighting the autocrine mode of action of FGF-2. At the endothelial membrane level, zofenoprilat appeared to activate the bradykinin B1 receptor, a known stimulant of FGF-2 expression. In conclusion, we show that ACEI exert protective/proangiogenic effects in microvascular coronary endothelial cells by activating the endogenous FGF-2/FGFR-1 system.
血管紧张素转换酶抑制剂（ACEI）对血管内皮产生的有益作用归因于内皮细胞存活特性的恢复和血管生成的改善。成纤维细胞生长因子（FGF）-2是微血管内皮的血管生成因子，它通过自分泌/旁分泌机制持续促进内皮细胞生长和存活。在此，我们提出假说，即FGF-2可能有助于ACEI的促存活/促血管生成作用。我们研究了佐芬普利拉，并在选定实验中研究了赖诺普利，作为ACEI的代表。这些化合物通过使内皮细胞生长增加高达5倍，诱导从猪冠状动脉和人脐动脉分离的血管片段中形成假毛细血管。一氧化氮合酶（NOS）途径抑制剂和抗FGF-2抗体可消除血管生成。同样，在培养的冠状动脉内皮细胞（CVEC）中，ACEI上调内皮型NOS（eNOS）和FGF-2，并诱导丝裂原活化蛋白激酶细胞外信号调节激酶1/2活化。eNOS/FGF-2的过表达在功能水平上产生了增强的细胞增殖和迁移，后一种作用具有剂量依赖性，在0.1μM佐芬普利拉时最大。用FGF-2的小干扰RNA转染CVEC后内皮功能受损，清楚地证明了FGF-2对ACEI引发的血管生成表型获得的重要性。此外，FGF-2沉默极大地影响了FGF受体（FGFR）-1的核转位，突出了FGF-2的自分泌作用模式。在内皮细胞膜水平，佐芬普利拉似乎激活了缓激肽B1受体，这是一种已知的FGF-2表达刺激物。总之，我们表明ACEI通过激活内源性FGF-2/FGFR-1系统在微血管冠状动脉内皮细胞中发挥保护/促血管生成作用。
